ORIGINAL ARTICLE Sequence analysis of the ribosomal DNA internal transcribed spacer regions in Termitomyces heimii species Shafiquzzaman Siddiquee & Weng Yan Yee & Khanam Taslima & Nur Hasan Nudin Fatihah & S. Vijay Kumar & Md Mainul Hasan Received: 29 October 2010 / Accepted: 8 July 2011 / Published online: 29 July 2011 # Springer-Verlag and the University of Milan 2011 Abstract Termitomyces represents a very poorly known genus of fungi whose essential characteristic is that all representatives of the genus are cultivated by termites (Macrotermitinae) in their nest. Many species of Termito- myces commonly form fruiting bodies which develop from fungus combs within the nest. Identification based on morphological characteristics is problematic, tedious, and prone to error. Therefore, correct identification at the species level is highly desirable. This is the first report on the identification of the Malaysian isolates of Termitomyces analyzed using the DNA sequence of their internal transcriber spacer regions (ITS1–5.8S–ITS2). The results clearly demonstrate that this group is clearly monophyletic and belongs to the Tricholomataceae family. Based on the morphological identification and the molecular analysis conducted in this study, we have determined that the isolates represent the termite fungi named Termitomyces heimii. Keyword Termitomyces heimii . Termites . Internal transcribed spacer (ITS) . Phylogenetic analysis Introduction The fungus-growing termites (Isoptera: Termitidae: Macro- termitinae) cultivate fungal symbionts of the monophyletic genus Termitomyces (Basidiomycota: Tricholomataceae: Termitomyceteae) on specially constructed substrate combs located in single or multiple nest chambers (Wood and Thomas 1989; Aanen et al. 2002; Frøslev et al. 2003). These combs consist of finely fragmented dried plant and wood material with a high C/N ratio. Once fungal degradation occurs, the termites use both the fungal tissues (mycelium and asexual fruiting bodies called nodules) and the degraded plant materials. The relative proportions of fungal and plant material in the termite diet change between castes and between species (Leuthold et al. 1989; Traniello and Leuthold 2000). Termites belonging to the subfamily Macrotermitinae cultivate the mycelium of basidiomycetes on comb-like structures made of small spheres of plant material which quickly passes through the termite gut. The fungi produce small nodules which are consumed by the termites along with the degraded comb. In this way, the exosymbiotic fungus makes complex plant macromolecules for the termites, parallel to the function of the protozoic endosymbionts of non-fungus growing termites (Rouland Lefevre 2000). During S. Siddiquee (*) : S. V. Kumar Biotechnology Research Institute, Universiti Malaysia Sabah JLN UMS, 88400 Kota Kinabalu, Sabah, Malaysia e-mail: shafiqpab@ums.edu.my W. Y. Yee Department of Biology, Faculty of Science, Universiti Putra Malaysia UPM, 43400 Serdang, Selangor, Malaysia K. Taslima Department of Fisheries Biology and Genetics, Faculty of Fisheries, Bangladesh Agricultural University, Mymensingh, Bangladesh N. H. N. Fatihah Department of Agricultural Science, Faculty of Agriculture & Biotechnology, Universiti Sultan Zainal Abidin (UNISZA), 20400 Kuala Terengganu, Malaysia M. M. Hasan Department of Agricultural Botany, Patuakhali Science and Technology University, Dumki, Patuakhali 8602, Bangladesh Ann Microbiol (2012) 62:797–803 DOI 10.1007/s13213-011-0320-8